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1. Protocols for Rearing FallWebworm (Hyphantria cunea) in a Colony and Basic Methods for Laboratory and Field Experiments

   Robinson-Castillo, M.; Mitchell, A.; Fasching, A.; Vidal, M. C.; Abarca, M.; Wimp, G. M.; Bieber, B.; Vyas, D. K.; Murphy, S.M.. (January 2021, University of Denver Undergraduate Research Journal)

   null (Ed.)

   Standardized protocols are an essential asset for research requiring the maintenance of live organisms. Ecological studies often involve collaborations between multiple teams that are spread across locations, and these collaborations benefit from sharing successful laboratory procedures. Our research team is studying the ecology of the fall webworm moth (Hyphantria cunea, hereafter FW) in North America for >10 years, during which time we have established reliable procedures for starting and maintaining FW colonies under laboratory conditions. FW is a North American species that has been introduced to Europe and Asia where it is a major pest. Here, we present a detailed review of the methods we use to find and collect FW caterpillars in the field, house and rear caterpillars in the laboratory, handle pupae, and initiate diapause for overwintering. We also describe how to end diapause the following summer, care for emerging adult moths and mate them, and tend to eggs. Lastly, we test the effectiveness of some of our protocols related to mating adult moths to determine whether fertile eggs are produced. FW is becoming a model study system for ecological and evolutionary studies related to diet breadth. As more researchers begin studying the ecology and management of FW, laboratory colonies will play an important role for these projects. Our protocols will provide guidance to inform the successful study of this important insect. 

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2. Protocols for rearing fall webworm (Hyphantria cunea) in a colony and basic methods for laboratory and experiments

   Robinson-Castillo, M. (January 2021, University of Denver Undergraduate Research Journal)

   null (Ed.)

   Standardized protocols are an essential asset for research requiring the maintenance of live organisms. Ecological studies often involve collaborations between multiple teams that are spread across locations, and these collaborations benefit from sharing successful laboratory procedures. Our research team has studied the ecology of the fall webworm moth (Hyphantria cunea, hereafter FW) in North America for >10 years, during which time we have established reliable procedures for starting and maintaining FW colonies under laboratory conditions. Here, we present a detailed review of the methods used to find and collect FW caterpillars in the field, house and rear caterpillars in the laboratory, handle pupae, and initiate diapause for overwintering. We also describe how to end diapause the following summer, care for emerging adult moths and mate them, and tend to eggs. Lastly, we test the effectiveness of some of our protocols related to mating adult moths to determine whether fertile eggs are produced. FW is a North American species that has been introduced to Europe and Asia where it is a major pest. FW is also becoming a model study system for ecological and evolutionary studies related to diet breadth. As more researchers begin studying the ecology and management of FW, laboratory colonies will play an important role for these projects. Our protocols will provide guidance to inform the successful study of this important insect. 

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3. Exposure to Non-Native Tropical Milkweed Promotes Reproductive Development in Migratory Monarch Butterflies

   https://doi.org/10.3390/insects10080253  

   Majewska, Ania A.; Altizer, Sonia (August 2019, Insects)

   Background: North American monarchs (Danaus plexippus) are well-known for their long-distance migrations; however, some monarchs within the migratory range have adopted a resident lifestyle and breed year-round at sites where tropical milkweed (Asclepias curassavica) is planted in the southern coastal United States. An important question is whether exposure to exotic milkweed alters monarch migratory physiology, particularly the ability to enter and remain in the hormonally-induced state of reproductive diapause, whereby adults delay reproductive maturity. Cued by cooler temperatures and shorter photoperiods, diapause is a component of the monarch’s migratory syndrome that includes directional flight behavior, lipid accumulation, and the exceptional longevity of the migratory generation. Methods: Here, we experimentally test how exposure to tropical milkweed during the larval and adult stages influences monarch reproductive status during fall migration. Caterpillars reared under fall-like conditions were fed tropical versus native milkweed diets, and wild adult migrants were placed in outdoor flight cages with tropical milkweed, native milkweed, or no milkweed. Results: We found that monarchs exposed to tropical milkweed as larvae were more likely to be reproductively active (exhibit mating behavior in males and develop mature eggs in females) compared to monarchs exposed to native milkweed. Among wild-caught fall migrants, females exposed to tropical milkweed showed greater egg development than females exposed to native or no milkweed, although a similar response was not observed for males. Conclusions: Our study provides evidence that exposure to tropical milkweed can increase monarch reproductive activity, which could promote continued residency at year-round breeding sites and decrease monarch migratory propensity. 

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4. Points to Consider When Establishing and Rearing Culex Mosquitoes in the Laboratory

   https://doi.org/10.1101/pdb.top107823  

   Meuti, Megan E; Siperstein, Alden; Wolkoff, Matthew (August 2023, Cold Spring Harbor Protocols)

   Culexmosquitoes transmit several pathogens to humans and animals, including viruses that cause West Nile fever and St. Louis encephalitis and filarial nematodes that cause canine heartworm and elephantiasis. Additionally, these mosquitoes have a cosmopolitan distribution and provide interesting models for understanding population genetics, overwintering dormancy, disease transmission, and other important and ecological questions. However, unlikeAedesmosquitoes that produce eggs that can be stored for weeks at a time, no obvious “stopping” point exists in the development ofCulexmosquitoes. Therefore, these mosquitoes require nearly continuous care and attention. Here, we describe some general considerations when rearing laboratory colonies ofCulexmosquitoes. We highlight different methods so that readers may choose what works best for their experimental needs and laboratory infrastructure. We hope that this information will enable additional scientists to conduct laboratory research on these important disease vectors. 

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5. Rearing and Shipping of Uranotaenia lowii, a Frog-Biting Mosquito

   https://doi.org/10.21769/BioProtoc.4996  

   Singh, Richa; Sanscrainte, Neil; Estep, Alden; González, Katherine; Bernal, Ximena E (June 2024, BIO-PROTOCOL)

   Many studies on mosquito biology rely on laboratory-reared colonies, emphasizing the need for standardized protocols to investigate critical aspects such as disease biology, mosquito behavior, and vector control methods. While much knowledge is derived from anthropophilic species from genera like Anopheles, Aedes, and Culex, there is a growing interest in studying mosquitoes that feed on non-human hosts. This interest stems from the desire to gain a deeper understanding of the evolution of diverse host range use and host specificity. However, there is currently a limited number of comprehensive protocols for studying such species. Considering this gap, we present a protocol for rearing Uranotaenia lowii, a mosquito species specialized in feeding on anuran amphibians by eavesdropping on host-emitted sound cues. Additionally, we provide instructions for successfully shipping live specimens to promote research on this species and similar ones. This protocol helps fill the current gap in comprehensive guidelines for rearing and maintaining colonies of anuran host–biting mosquitoes. It serves as a valuable resource for researchers seeking to establish colonies of mosquito species from the Uranotaeniini tribe. Ultimately, this protocol may facilitate research on the evolutionary ecology of Culicidae, as this family has recently been proposed to have originated from a frog-feeding ancestor. 

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